No abstract available.
Paramyxoviridae Infections* ; Polymerase Chain Reaction* ; Reverse Transcription*

Laccase activity of Pleurotus ostreatus is significantly increased by the addition of apple pomace. Among various conditions, the best concentration of apple pomace and cultivation time for the production of laccase by P. ostreatus was 2.5% and 9 days, respectively. Reverse transcription polymerase chain reaction analyses of laccase isoenzyme genes, including pox1, pox3, pox4, poxc, poxa3, and poxa1b, revealed a clear effect of apple pomace on transcription induction. Our findings reveal that the use of apple pomace can be a model for the valuable addition of similar wastes and for the development of a solid-state fermenter and commercial production of oyster mushroom P. ostreatus.
Fungi* ; Laccase* ; Pleurotus* ; Polymerase Chain Reaction ; Reverse Transcription

Objective. To determine the diagnostic accuracy of self-collected snorted and spit saliva in detecting COVID-19 using RT-PCR (ssRT-PCR) and lateral flow antigen test (ssLFA) versus nasopharyngeal swab RT-PCR (npRT-PCR). Methods. One hundred ninety-seven symptomatic subjects for COVID-19 testing in a tertiary hospital underwent snort-spit saliva self-collection for RT-PCR and antigen testing and nasopharyngeal swab for RT-PCR as reference. Positivity rates, agreement, sensitivity, specificity, and likelihood ratios were estimated. Results. Estimated prevalence of COVID-19 using npRT-PCR was 9% (exact 95% CI of 5.5% - 14.1%). A higher positivity rate of 13% in the ssRT-PCR assay suggested possible higher viral RNA in the snort-spit samples. There was 92.9% agreement between ssRT-PCR and npRT-PCR (exact 95% CI of 88.4% to 96.1%; Cohen’s Kappa of 0.6435). If npRT-PCR will be assumed as reference standard, the estimated Sensitivity was 83.3% (exact 95% CI of 60.8% to 94.2%), Specificity 93.9% (exact 95% CI of 89.3% to 96.5%), Positive predictive value of 57.7% (exact 95% CI of 38.9% to 74.5%), Negative predictive value of 98.2% (exact 95% CI of 95% to 99.4%), positive likelihood ratio of 3.65 (95% CI of 7.37 to 24.9), negative likelihood ratio of 0.178 (95% CI of 0.063 to 0.499). There was 84.84% agreement (95% exact CI of 79.1% to 89.5%; Cohen’s Kappa of 0.2356) between ssLFAvs npRT-PCR, sensitivity of 38.9% (exact 95% CI of 20.3% to 61.4%), specificity of 89.4% (exact 95% CI of 84.1% to 93.1%), PPV of 26.9% (95% CI of 13.7% to 46.1%), NPV of 93.6% (exact 95% CI of 88.8% to 96.4%), LR+ of 3.67 (95% CI of 1.79 - 7.51), LR – of 0.68 (95% CI of 0.47 - 0.99). Conclusion. Our data showed that snort-spit saliva RT-PCR testing had acceptable diagnostic performance characteristics and can potentially be used as an alternative to the standard nasopharyngeal/oropharyngeal swab RT-PCR test for COVID-19 in certain situations. However, our data also showed that snort-spit saliva antigen testing using lateral flow assay did not offer acceptable performance.
Saliva ; SARS-CoV-2 ; Reverse Transcription ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUND: Kruppel-like factor 4 (KLF4) is a transcription factor that regulates a diverse array of cellular processes, including development, differentiation, proliferation, and apoptosis. Although its function in keratinocytes has been widely studied, its exact role in psoriasis has not been elucidated. OBJECTIVE: We designed this study to investigate epidermal expression levels of KLF4 and the change in KLF4 expression after treatment in patients with psoriasis. METHODS: We compared the expression levels of KLF4 in the basal, suprabasal, and superficial epidermal layers, in psoriatic lesional, non-lesional, and normal skin, using an immunoreactivity intensity distribution index (IRIDI). In addition, we measured the change in KLF4 expression on the basis of the IRIDI and by reverse transcription polymerase chain reaction (RT-PCR) analysis after treatment. RESULTS: The combined IRIDI scores in psoriatic lesional skin were significantly higher than the scores in both non-lesional and normal skin. The psoriatic epidermis, particularly the suprabasal layer, showed a significantly increased IRIDI score compared to that of non-lesional and normal skin, which was significantly decreased after treatment. RT-PCR analysis exhibited a slight increase in KLF4 mRNA expression level after treatment; however, this increase was not significant. CONCLUSION: These data indicate that KLF4 could regulate epidermal proliferation and differentiation. Moreover, we believe that KLF4 may play an important role in the physiological reaction to counteract abnormal differentiation and proliferation of keratinocytes.
Apoptosis ; Epidermis ; Humans ; Keratinocytes ; Polymerase Chain Reaction ; Psoriasis* ; Reverse Transcription ; RNA, Messenger ; Skin ; Transcription Factors

This study was purposed to investigate the expression of the growth-factor independence 1 (GFI1) in patients with leukemia and its clinical significance. Bone marrow mononuclear cells were obtained from 65 newly diagnosed leukemia patients including 24 acute myeloid leukemia (AML), 18 chronic myelogenous leukemia (CML), 6 acute lymphoblastic leukemia (ALL), 17 blast crisis of chronic myelogenous leukemia and 13 patients with iron deficiency anemia (IDA) were used as controls. The relative expression of gene gfi1 was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and taqman quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR). The results showed that gene expression of gene gfi1 in leukemia patients was obviously higher than that in controls and the difference was statistically significant (p < 0.01), in which the expression of gene gfi1 in newly diagnosed CML patients was higher than that in newly diagnosed AML, newly diagnosed ALL, CML-BCP patients and the difference was significant (p < 0.01). Expression of gene gfi1 in lymphocytic blast crisis of CML was higher than that in nonlymphocytic blast crisis of CML, and the difference was significant. It is concluded that gene gfi1 may play an important role in leukemia, especially in CML incidence and progression. The high level expression of gene gfi1 may be participate in the development of lymphoma.
DNA-Binding Proteins ; genetics ; Gene Expression ; Humans ; Leukemia ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics ; Transcription, Genetic ; genetics

BACKGROUND: The detection of antibody to hepatitis C virus (anti-HCV) is the most useful method to investigate past or current HCV infections. We performed a clinical evaluation of ADVIA Centaur HCV assay, a new third generation assay for the qualitative detection of IgG antibody. METHODS: Included in the study were a total of 323 samples (108 positive and 215 negative), for which HCV reverse transcription polymerase chain reaction (RT-PCR) was requested. ADVIA Centaur HCV assay (Bayer Healthcare LLC, Diagnostics Division, Tarrytown, NY, USA) was compared with a currently available and widely used AxSYM HCV version 3.0 assay (Abbott Laboratories, Abbott Park, IL, USA). Samples with discrepant results were retested with each assay, and further tested with a recombinant immunoblot assay (RIBA, LG HCD Confirm, LG Chemical Co., Seoul, Korea). Reproducibility of Centaur assay was evaluated in five groups (two samples in each group) with different index values. RESULTS: The overall concordance rate was 91.6% (296/323) between Centaur and AxSYM assays. It was 100% (108/108) in RT-PCR positive samples and 87.4% (188/215) in RT-PCR negative samples. Discrepant samples (8.4%, 27/323) were all RT-PCR negative, and all except two were Centaur negative and AxSYM positive. In discrepant samples, RIBA showed negative results except for two samples with indeterminate results. The sensitivity and specificity of Cenyaur assay were 98.1% and 65.6%, and the respective figures for AxSYM assay were 98.1% and 54.9%. Reproducibility of Centaur assay was satisfactory. CONCLUSIONS: The overall concordance between Centaur and AxSYM assays was satisfactory. Sensitivity and specificity of Centaur assay were equivalent to or better than those of AxSYM assay. ADVIA Centaur HCV assay seems to be a reliable and useful method for the detection of anti-HCV in clinical laboratories.
Delivery of Health Care ; Hepacivirus* ; Immunoglobulin G ; Polymerase Chain Reaction ; Reverse Transcription ; Sensitivity and Specificity ; Seoul

PURPOSE: The objective of this study was to investigate the effects of (6)-gingerol, ginger components proliferation and adipocyte differentiation from early to lately steps. METHODS: 3T3-L1 preadipocytes were cultured. Differentiation of confluent cells was induced with dexamethasone, isobutylxanthin and insulin for 2 day and cells were cultured by medium with insulin in presence of various concentrations 0, 25, 50, 100 (micromol/L) of (6)-gingerol for 4 day. Cell viability was measured using the EZ Cytox assay kit. In addition, we examined the expression of mRNA levels associated with each adipocyte differentiation step by real time reverse transcription polymerase chain reaction. RESULTS: (6)-Gingerol inhibited adipocyte proliferation in a dose and time dependent manner. Expression of C/EBPbeta, associated with early differentiation step remained unchaged. However, intermmediate, late differentiation step and adipocytokines were effectively changed in dose-dependently manner in cell groups treated with (6)-gingerol. CONCLUSION: This study has shown that treatment with (6)-gingerol inhibited adipocyte proliferation as well as each adipocyte differentiation step. In particular, the (6)-gingerol more effectively inhibited adipocyte differentiation from intermmediate differentiation step.
Adipocytes* ; Adipokines ; Cell Survival ; Dexamethasone ; Ginger* ; Insulin ; Polymerase Chain Reaction ; Reverse Transcription ; RNA, Messenger

OBJECTIVE: To overcome the limitations of the single gene transfer, the authors present the results of wild-type p16 and p53 combined genes transfer in vitro to the U251MG and U373MG cell lines using cationic liposome as a vector. METHODS: To compare the therapeutic effect of the combined p16 and p53 genes transfer with the single p16 and p53 gene transfer, full length of wild-type human p16 and p53 gene, and combined p16-p53 genes were transferred in vitro to the U251MG and U373MG cell lines using cationic liposome as a vector. As the U251MG and U373MG cell lines are devoid of p16 and p53 genes, the therapeutic effect of the three groups of gene transfer could be evaluated by the growth suppression or percentage of the viable cells. Reverse transcription polymerase chain reaction(RT-PCR), flow cytometry, and electron microscopy(EM) were used for evaluation of the growth suppression or apoptosis of the tumor cells. RESULTS: p16 gene, p53 gene and the combined p16-p53 genes were effectively transferred to the cell lines using cationic liposome as a vector resulting in dramatic decrease of the viable tumor cells in comparison to the control group(p=0.004). The cytotoxic effect of the gene transfer in the U251MG cell line was the most significant in the combined p16-p53 group. However, in the U373MG cell line p53 single gene transfer group showed more significant effect than the combined gene transfer group. Apoptosis was confirmed by EM in the combined p16-p53 genes group. The G1 phase arrest effect, confirmed by the flow cytometry was more prevalent in the p16 gene transfer group than the other groups. CONCLUSION: Cationic liposome-mediated transfer of combined p16-p53 genes to the human glioblastoma cell lines is proven effective. However, the therapeutic effect of the combined p16-p53 genes transfer was not consistently superior to the single p16 or p53 gene transfer.
Apoptosis ; Cell Line* ; Flow Cytometry ; G1 Phase ; Genes, p16 ; Genes, p53* ; Glioblastoma* ; Humans* ; Liposomes ; Reverse Transcription

BACKGROUND: Norovirus is a leading cause of epidemic and sporadic acute gastroenteritis worldwide. Because of the rapid transmission of the virus, early detection is important to prevent outbreak of norovirus infection. To evaluate the performance of a newly introduced rapid antigen test for detecting human norovirus in stool specimens, we compared it with the established ELISA test and real time quantitative reverse transcription PCR (qRT-PCR). METHODS: One hundred and eighty-four stool samples were analyzed by rapid antigen test (Denka-Seiken, Japan), ELISA (R-Biopharm, Germany), and qRT-PCR (R-Biopharm, Germany). Overall percent agreement, percent positive agreement (PPA), and percent negative agreement (NPA) of the rapid antigen test in comparison with ELISA and qRT-PCR were obtained. RESULTS: Positive rates of rapid antigen test, ELISA, and qRT-PCR were 44.0% (81/184), 51.6% (95/184), and 42.9% (79/184), respectively. Seventy samples (38.0%) showed all positive, and 86 samples (46.7%) showed all negative results by three methods. Overall percent agreement of three methods was 84.8% (156/184). Overall percent agreement, PPA, and NPA of the rapid antigen test in comparison with qRT-PCR were 89.1%, 88.6%, and 89.5%, respectively, and those of the rapid antigen test in comparison with ELISA were 90.2%, 83.2%, and 97.8%, respectively. Total procedure of the rapid antigen test was finished within 20 min. CONCLUSIONS: Rapid antigen test was easier and quicker to perform, and showed high agreement rates with ELISA and qRT-PCR. This test may be useful for rapid screening of norovirus infection.
Enzyme-Linked Immunosorbent Assay ; Gastroenteritis ; Humans ; Mass Screening ; Norovirus ; Polymerase Chain Reaction ; Reverse Transcription ; Viruses

PURPOSE: Recently, two alternatively spliced survivin variants, survivin-DeltaEx3 and survivin-2B, were identified in a single copy of the survivin gene. It has been reported that the expressions of survivin splice variants significantly correlates with the clinical results in many types of human carcinoma. We investigated the transcription levels of survivin and its splice variants in human colorectal carcinomas, and analyzed correlations between survivin expression levels and clinicopathologic features. METHODS: We used Western blot and real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) to analyze the protein and mRNA expression levels of survivin variants in 51 colorectal carcinomas. The quantitative RT-PCR was performed using primer pairs specific for survivin and each of its splice variants, then normalized for the gene that encodes glyceraldehydes-3-phosphate dehydrogenase. RESULTS: In Western blotting, the protein levels of survivin were higher in the tumor tissue than in normal tissue. The expression of survivin, survivin-2B and survivin-DeltaEx3 mRNA was present in 96%, 64.7%, and 82.4% of the samples, respectively. When the pathologic parameters were compared, colorectal cancers of advanced pT stages showed significant decrease in survivin-2B mRNA expression by the quantitative RT-PCR (P < 0.001). CONCLUSION: The decreased expression of survivin-2B might be related to tumor progression in colorectal cancers. This finding indicates that alternatively spliced variants of survivin may be involved in refining the functions of survivin during tumor progression.
Blotting, Western ; Coat Protein Complex I ; Colorectal Neoplasms ; Humans ; Polymerase Chain Reaction ; Reverse Transcription ; RNA, Messenger